Comparative study for chiral separation of Atracurium besylate isomers: Eco-friendly HPLC using Lux Cellulose-3 chiral column, and TLC-Densitomerty approaches

Abstract

Liquid chromatography offers an efficient strategy for chiral drug separation using specific stationary phases or mobile solvent additives. Two approaches are performed for Atracurium besylate (ATR) chiral separation involving three isomers (cis-cis, cis-trans, trans-trans isomers). The initial method depended on reversed phase High Performance Liquid Chromatography (HPLC) employing Lux Cellulose-3 as a chiral stationary phase (CSP) and acetonitrile: water (90:10, v/v) as a mobile phase at a flow rate of 0.7 mL min−1 with UV detection at 280.0 nm. The obtained peaks of ATR isomers were fairly separated with excellent resolution values. The isomer ratio was found to be 57.89% cis-cis, 36.57% cis-trans, 5.54% trans-trans (expected ratio 55.0–60.0%: 34.5–38.5%: 5.00–6.50%, respectively). Linearity, precision and accuracy were proven to be acceptable over the concentration range of (10.0–100.0 μg mL−1) ATR. Utilizing CSP provided the best separation within short analysis time (3 min) and decreased the waste generation when compared to other reported methods. The alternative method was based on thin layer chromatography (TLC) chiral separation using β-cyclodextrin sulphated sodium salt (S-β-CD) as a chiral mobile phase additive. The separation was achieved on silica gel plates using acetonitrile: methanol: water: ethyl acetate: 1% acetic acid (5:2:1.5:1:0.5, by volume) as a developing system. The linear regression analysis data were assessed for the regression line in the range (1.0–15.0 μg band−1) of racemic ATR. The two proposed methods have been validated for ATR chiral separation in commercial vials, following the relevant FDA and ICH guidelines. The obtained results were statistically compared with those of the official HPLC method showing that there was no significant difference.